Comparative phenotype analysis for assessment of biological active compounds such as antimicrobials

Abstract

The present invention relates to using multitest panels to improve the effectiveness, throughput, and efficiency of testing and commercial development of biologically active compounds, in particular those useful in human, animal, and plant health. In particular, the present invention provides phenotype microarrays suitable for testing biologically active compounds for their potential application in clinical, veterinary, and plant health.

What is claimed is:  
     
       1. A method for detecting the combined effect of at least two biologically active chemicals in vivo, comprising the steps of: 
       a) providing:  
       i) a suspension comprising at least one cell and a first biologically active chemical, and  
       ii) a testing device having at least two wells, wherein at least one well further comprises at least one subsequent biologically active chemical, and wherein said first and said subsequent biologically active chemicals are different compounds;  
       b) inoculating said suspension into said wells of said testing device; and  
       c) detecting a cellular response of said at least one cell in said wells.  
     
     
       2. The method of  claim 1 , wherein said at least one cell is selected from the group consisting of a bacterial cell, a fungal cell, a mammalian cell, an insect cell and a plant cell. 
     
     
       3. The method of  claim 1 , wherein said at least two wells further comprise medium selected from the group consisting of microbiological medium and cell culture medium. 
     
     
       4. The method of  claim 1 , wherein said testing device comprises a plurality of wells and wherein said subsequent biologically active chemical is present in said wells at different concentrations. 
     
     
       5. The method of  claim 1 , wherein said detecting is with a colorimetric indicator in said suspension or in said testing device. 
     
     
       6. The method of  claim 5 , wherein said colorimetric indicator is selected from the group consisting of a chromogenic substrate, a fluorogenic compound, a luminogenic compound, a redox indicator and a pH indicator. 
     
     
       7. The method of  claim 6 , wherein said cellular response of said at least one cell is respiration, and wherein said redox indicator is a tetrazolium salt. 
     
     
       8. The method of  claim 1 , wherein said suspension further comprises a gelling agent, and wherein said testing device further comprises a gel-initiating agent in said wells. 
     
     
       9. The method of  claim 1 , wherein said detecting is done at a fixed time point, at frequent time intervals, or continuously. 
     
     
       10. The method of  claim 1 , further comprising step (d) comparing said cellular response of said at least one cell in the presence of said subsequent biologically active chemical to that of said at least one cell in the absence of said subsequent biologically active chemical. 
     
     
       11. The method of  claim 10 , further comprising step (e) recording the results of said comparing to generate a database of said cellular responses to said biologically active chemicals. 
     
     
       12. A method for examining the effects of at least two biologically active chemicals in vivo, comprising the steps of: 
       a) obtaining a metabolic profile for a first biologically active chemical by detecting a response of a cell suspension incubated in the presence of a first biologically active chemical in a testing device comprising a plurality of wells each containing a different substrate;  
       b) obtaining a metabolic profile for at least one subsequent biologically active chemical by detecting a response of a cell suspension incubated in the presence of a subsequent biologically active chemical in a testing device comprising a plurality of wells each containing a different substrate; and  
       c) creating a database of said metabolic profiles.  
     
     
       13. The method of  claim 12 , wherein said cell suspension is selected from the group consisting of a bacterial cell suspension, a fungal cell suspension, a mammalian cell suspension, an insect cell suspension, and a plant cell suspension. 
     
     
       14. The method of  claim 12 , wherein said detecting is with a colorimetric indicator selected from the group consisting of a chromogenic substrate, a fluorogenic compound, a luminogenic compound, a redox indicator and a pH indicator. 
     
     
       15. The method of  claim 14 , wherein said response of said cell suspension is respiration, and wherein said redox indicator is a tetrazolium salt. 
     
     
       16. The method of  claim 12 , wherein said detecting is done at a fixed time point, at frequent time intervals, or continuously. 
     
     
       17. The method of  claim 12 , wherein said at least two biologically active chemicals comprise at least 20 biologically active chemicals. 
     
     
       18. The method of  claim 12 , wherein said at least one subsequentbiologically active chemical comprises a drug candidate. 
     
     
       19. The method of  claim 12 , further comprising step (d) grouping said biologically active chemicals according to the similarity of said metabolic profiles in said database. 
     
     
       20. The method of  claim 19 , wherein said grouping comprises creating a cluster diagram. 
     
     
       21. The method of  claim 20 , wherein said cluster diagram comprises a dendrogram. 
     
     
       22. The method of  claim 19 , further comprising steps: 
       (e) obtaining a metabolic profile for a drug candidate by detecting a response of a cell suspension incubated in the presence of a drug candidate in a testing device comprising a plurality of wells each containing a different substrate; and  
       (f) determining whether said drug candidate falls into a group defined in step (d).